| 日 時 | 2025年02月27日(木) 15:00 より 16:00 まで |
|---|---|
| 講演者 | Prof Keisuke Ohta |
| 講演者所属 | Professor, Kurume University, School of Medicine, Fukuoka |
| 場 所 | Zoom オンライン |
| お問い合わせ先 | 窪田芳之 (yoshiy@nips.ac.jp) |
| 要旨 |
Volume electron microscopy (vEM) has revolutionised three-dimensional ultrastructural analysis, allowing the visualisation of previously hidden mesoscale biological structures. However, a fundamental limitation of electron microscopy is the difficulty in correlating ultrastructure with molecular identity. Unlike fluorescence microscopy, which uses fluorescent proteins and immunolabelling to link structural and biochemical information, conventional electron microscopy lacks intrinsic molecular contrast, making identifying cell types highly dependent on prior knowledge. Immuno-electron microscopy and correlative light and electron microscopy (CLEM) have been developed to address this gap. However, the need for electron microscopy compatible fixation often results in loss of antigenicity, and osmium tetroxide (OsO₄) quenching severely limits fluorescence retention. To overcome this problem, various CLEM methods have been developed, adapted to specific applications. Recently, the development of OsO₄-resistant fluorophores has enabled fluorescence retention even after electron microscopy-compatible fixation, resin embedding and ultrathin sectioning, and new age CLEM methods (in-resin CLEM) have been reported. Furthermore, if electron microscopy compatible fixation strategies can be achieved without glutaraldehyde - commonly used in pathological specimens - this could open the way for advanced CLEM workflows in pathology. Furthermore, the integration of tissue-based cell identification with vEM could significantly improve our ability to correlate molecular and ultrastructural data. In this talk, we will present our recent efforts to extend the applicability of in-resin CLEM to 3D ultrastructural imaging. |
